Abstract
Evidence is rapidly mounting that transposable element (TE) expression and replication may impact biology more widely than previously thought. This includes potential effects on normal physiology of somatic tissues and dysfunctional impacts in diseases associated with aging, such as cancer and neurodegeneration. Investigation of the biological impact of mobile elements in somatic cells will be greatly facilitated by the use of donor elements that are engineered to report de novo events in vivo. In multicellular organisms, reporter constructs demonstrating engineered long interspersed nuclear element (LINE-1; L1) mobilization have been in use for quite some time, and strategies similar to L1 retrotransposition reporter assays have been developed to report replication of Ty1 elements in yeast and mouse intracisternal A particle (IAP) long terminal repeat (LTR) retrotransposons in cultivated cells. We describe a novel approach termed cellular labeling of endogenous retrovirus replication (CLEVR), which reports replication of the gypsy element within specific cells in vivo in Drosophila. The gypsy-CLEVR reporter reveals gypsy replication both in cell culture and in individual neurons and glial cells of the aging adult fly. We also demonstrate that the gypsy-CLEVR replication rate is increased when the short interfering RNA (siRNA) silencing system is genetically disrupted. This CLEVR strategy makes use of universally conserved features of retroviruses and should be widely applicable to other LTR retrotransposons, endogenous retroviruses (ERVs), and exogenous retroviruses.
Highlights
50% of human DNA content, and equivalently vast fractions of most other animal and plant genomes, consist of sequences derived from transposable elements (TEs) [1]
We demonstrate that the gypsy-cellular labeling of endogenous retrovirus replication (CLEVR) construct reports gypsy replication events both in cell culture and in several cell types in the adult Drosophila central nervous system
We found that the activation of the gypsy-CLEVR reporter is age dependent in neurons and in glial cells
Summary
50% of human DNA content, and equivalently vast fractions of most other animal and plant genomes, consist of sequences derived from transposable elements (TEs) [1]. Selfish genetic elements whose primary adaptation is to copy themselves in germline tissue, thereby passing on de novo copies. Replication in the germline is the means by which TEs generate inheritable de novo copies, it is increasingly clear that endogenous TEs replicate in somatic tissues. The evidence for somatic transposition is strong for Class 1 TEs, known as retrotransposable elements (RTEs) [4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21]. Somatic replication of RTEs has the potential for tremendous impact both on normal biological properties of tissues and on human health [2, 16,17,18,19,20,21]
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